The proposed application represents a comprehensive plan of three projects to improve the capacity of existing tests for pancreatic cancer, and to validate novel biomarkers discovered as part of collaborative efforts during the previous phase of the EDRN. The first project further develops a broad based diagnostic test for pancreatic cancer that enhances the CA19-9 test and incorporates elements of early detection of cancer by detecting autoantibodies to tumor specific glycopeptide structures found on mucins. The earliest premalignant and malignant lesions in the pancreas express MUCI, MUC4, MUC5ac, MUC16, and MUC17 with truncated O-linked structures attached (Tn, sialyl Tn, T) but may not produce a sufficient amount to be detected in serum. Instead, autoantibodies to these specific structures are produced at sufficiently high concentrations to be detected. As lesions progress mucins are detected in circulation, initially as immune complexes with the autoantibodies, and in later stages in the absence of immune complexes. This overall hypothesis will be tested by developing a set of three integrated tests for serum: a test that quantifies autoantibodies, a test that quantifies immune complexes, and a test that quantifies circulating mucins for both oligosaccharide and core protein (on the same molecule). A complementary study in collaboration with Dr. Sam Hanash will identify non-mucin proteins that carry Tn and sialyl Tn antigens and evaluate autoantibodies to these proteins. The second project led by Dr. Surinder Batra proposes to further develop a novel analytical test that shows enhanced sensitivity for detection mucin type glycoprotein (which can be difficult to detect using conventional techniques) using surface enhanced Raman spectroscopy (SERS). This technique will be used to develop an assay for multiple biomarkers in serum. The third project, led by Dr. Schmittgen and in collaboration with Dr. Hollingsworth, will undertake discovery of potential miRNA biomarkers in plasma and urine of patients with different stages of pancreatic cancer and evaluate the prognostic utility for patients with resected pancreatic cancer of evaluating the tissue expression of a set of microRNAs that are associated with early metastasis of pancreatic cancer.